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156-161 Research Article Evaluation of Antibacterial Activity Of Available online www.jocpr.com Journal of Chemical and Pharmaceutical Research, 2014, 6(8):156-161 ISSN : 0975-7384 Research Article CODEN(USA) : JCPRC5 Evaluation of antibacterial activity of extracts from Calendula aventis flowers Abdul-Malik Abudunia*1,2,4 , M’Hammed Ansar 2, Jamal Taoufik 2, Youssef Ramli 3, El Mokhtar Essassi 3, Azeddine Ibrahimi 1 and Khadija Khedid 4 1Biotechnology Laboratory (Med-Biotech), Faculty of Medicine and Pharmacy, Mohammed V University-Souissi, Rabat, Morocco 2Department of Medicinal Chemistry Laboratory, Faculty of Medicine and Pharmacy, Mohammed V University- Rabat, Morocco 3Department of Heterocyclic Organic Chemistry Laboratory, Faculty of Sciences, Mohammed V University-Agdal, Rabat, Morocco 4Department of Bacteriology, National Institute of Health (INH), Avenue, Ibn Batouta -Agdal, Rabat, Morocco ____________________________________________________________________________________________ ABSTRACT Calendula arvensis (C.A.) is one of the herbs used in traditional medicine in Morocco , Due to its essential oil it is recognized by its smell and warmth characteristic .This work aims to evaluate the antibacterial activity of C.A. flower extracts , Hexanolic and methanolic extracts were obtained by the method of soxhlet , while aqueous extract was obtained by maceration in cold water. Antibacterial activity screening was conducted by the well diffusion method according to the clinical and laboratory standard institute guidelines using 6 Gram positive and 7 Gram negative bacteria (sensitive and multidrug-resistant) .The results indicated that , the methanolic extracts of C.A. flowers generally showed high inhibitory activity against Gram-positive and Gram negative bacteria except Acinetobacter baumanii , Proteus mirabilis and Listeria monocytogenes, The C.A. flowers hexanolic extract inhibited all bacteria of Gram-positive and Gram negative bacteria except, Staphylococcus aureus MRSA, Streptococcus agalactiae and Acinetobacter baumanii , There were no inhibitory effects of the aqueous extracts against all tested bacterial strains except Rhodococcuss equi and Morganella morganii, MICs values of methanolic extracts were between 12.5-25 µg/ml . while MICs of values of hexanolic extracts were between 6.25- 12.5 µg/ml . In conclusion , the obtained results show an antibacterial potency of organic extracts of C.A. flowers. C.A. flowers Hexanolic extracts were bactericidal for most bacteria while methanolic extracts were bacteriostatic . Key words: Calendula arvensis , antibacterial activity, MICs, MBCs, medicinal plant. ____________________________________________________________________________________________ INTRODUCTION In Morrocco , as elsewhere in the world, plants are widely used in the treatment of various diseases . The varied climate and heterogeneous ecologic condition in Morocco have favoured the proliferation of more than 42,000 species of plants, divided into 150 families and 940 genuses [1]. Drugs derived from natural sources play a significant role in the prevention and treatment of human diseases. In many developing countries, traditional medicine is one of the primary healthcare systems [2,3]. Herbs are widely exploited in the traditional medicine and their curative potentials are well documented [ 4]. Therefore, in the traditional medicine prevention and treatments of several humans diseases is succefully controlled by using aromatics medicinal plants . Recently, in pharmaceutical industry, about 61% of new drugs developed between 1981 and 2002 derived from natural sources and they have been very successful, especially in the areas of infectious disease and cancer [ 5].Natural products of higher plants may give a new source of antimicrobial agents with possibly novel mechanisms of action [ 6,7]. The effects of plant extracts on bacteria have been studied by a very large number of researchers in different parts of the world [ 8] . Plants are rich in a wide variety of secondary metabolites such as tannins, terpenoids, alkaloids, flavonoids, 156 Abdul-Malik Abudunia et al J. Chem. Pharm. Res., 2014, 6(8):156-161 ______________________________________________________________________________ glycosides, etc., which have been found in vitro to have antimicrobial properties [ 9,10 ]. Herbal medicines have been known to man for centuries. Therapeutic efficacy of many indigenous plants for several disorders has been described by practitioners of traditional medicine [ 11 ]. Antimicrobial properties of medicinal plants are being increasingly reported from different parts of the world. The World Health Organization estimates that plant extracts or their active constituents are used as folk medicine in traditional therapies of 80% of the world's population [ 12 ]. The harmful microorganisms can be controlled with drugs and these results in the emergence of multiple drug-resistant bacteria and it has created alarming clinical situations in the treatment of infections. The pharmacological industries have produced a number of new antibiotics resistance to these drugs by microorganisms has increased. In general, bacteria have the genetic ability to transmit and acquire resistance to synthetic drugs which are utilized as therapeutic agents [ 13 ]. Calendula L . ( Asteraceae ), usually known as “marigold ”, is a reputed medicinal plant with ornamental properties. The yellow or orange-colored flowers are used as food dye, spice, and tea as well as tincture, ointment or cosmetic cream. Although the genus Calendula is usually indigenous to the southern European region including Italy, Malta, Greece, Turkey, Portugal, and Spain [14], it is nowadays cultivated in many temperate regions of the world depending on its commercial value. Since Calendula officinalis is grown in northern parts of Africa, it is also named as “African marigold ” [15]. Calendula is a relatively small genus (about 12–20 species native to the Macaronesian and Mediterranean regions are currently recognized) that belongs to the Asteraceae family (tribe: Calendula ) [16] . Calendula arvensis C.A. the common name of which is field marigold, is an annual herbaceous species (10–100 cm tall) that is widespread in central and southern Europe, northern Africa, southwestern Asia and the Macaronesian region [17] . The leaves, which contain tector and secretory trichomes, are lance shaped. The inflorescence consists of a single flower head with yellow or orange capitula (blossoms are produced year round, but flowering peaks between March and July). From a taxonomic viewpoint, C.A. is a complex species because of polymorphism in the size of the stems (5–60 cm), width of the leaves (5–20 mm), color of the flowers (bright yellow to yellow-orange) and the size and shape of the three types of achene (rostrate , cymbiform and annular) [18]. Thus, various subspecies have been ascribed to C.A. in various Mediterranean areas [16]. Numerous investigations have proved that extracts of the aerial parts of C.A. are rich in sesquiterpene glycosides (based on their alloaromadendrane, eudesmane and cubebane skeletons) [19]. Furthermore, phytochemical studies have shown that C.A. contains various families of compounds such as the phenolic acids and the flavonoids. To our knowledge, no data are available on the chemical composition of essential oil obtained from C.A. Several investigations on the volatile constituents of Calendula officinalis (also known as the pot marigold) have been conducted in which volatiles were extracted by hydrodistillation [20]. Calendula officinalis oils are generally characterized by a high content of sesquiterpenes with cadinane or muurolane skeletons. It should be noted that this species is cultivated worldwide for ornamental and medicinal purposes. It is also used in the manufacture of cosmetics, perfumes and pharmaceuticals and in the food industry to add color or flavor. There is only one report on the chemical composition of the essential oils of other Calendula species, viz., a report on the essential oil composition of the flowers of Calendula micrantha plants in Egypt [21] . The main components were aromadendrene, a-gurjunene, a- pinene, benzyl formate, (E)-b-caryophyllene and guaiol [22]. The aim of this study is to evaluate the antibacterial activity of C.A. flowers of methanolic extracts, hexanolic and aqueous extracts, and to, therefore, determine the scientific basis for its use in traditional medicine in the treatment of infection diseases. To the best of our knowledge, this is the first study to do so against a wide range of bacteria. EXPERIMENTAL SECTION Plant Materials: Flowers of Calendula arvensis were collected based on ethno pharmacological information from the villages around the region Rabat-Temara -Sidi El-Abed, during the month of March 2011 starting around 7 AM each morning. All agreements were obtained from the authorities with respect to the United Nations Convention of Biodiversity and with assistance of traditional medical practitioner. The plant was identified by botanist From the Moroccan Scientific Institute (Rabat). Avoucher specimen (N°RAB 78161) was deposited at the Herbarium of the Botany Department at the same institution. Preparation of the extracts : Aqueous Extract (AE) : The powdered of C.A. flowers extracts (200g)were extracted with boiling water (200 ml) for 30 min . The decoction was filtered and then freeze-dried (aqueous extract) [23]. Organic Extracts (OE) : Hexanolic extract (H.E.) and Methanolic extract (M.E.) were obtained by using the Soxhlet extraction of 200 g of C.A. flowers
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